Activation of the Mitogen Activated Protein Kinase Extracellular Signal-Regulated Kinase 1 and 2 by the Nitric Oxide–cGMP–cGMP-Dependent Protein Kinase Axis Regulates the Expression of Matrix Metalloproteinase 13 in Vascular Endothelial Cells

Abstract

Matrix metalloproteinases (MMPs) are synthesized in response to diverse stimuli, including cytokines, growth factors, hormones, and oxidative stress. Here we show that the nitric oxide (NO) donor 2-(N,N-diethylamino)-diazenolate-2-oxide (DEA-NO) and NO from murine macrophages transcriptionally regulate MMP-13 expression in vascular endothelial cells (BAEC). The cGMP analog, 8-bromo-cGMP (8-Br-cGMP) mimicked the effect of NO, whereas incubation with the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, or the cGMP-dependent protein kinase (PKG) inhibitor phenyl-1,N 2 -etheno-8-bromoguanosine-3Ј,5Ј-cyclic monophosphorothioate, Rp-isomer (PET) reduced the stimulatory effect of DEA-NO on the activation of the MMP-13 promoter. Overexpression of the catalytic subunit of PKG1-␣ resulted in a 5-to 6-fold increase of the MMP-13 regulatory region over control cells. On the other hand, incubation with the mitogen-activated protein/ extracellular signal-regulated kinase inhibitor 2Ј-amino-3Ј-methoxyflavone (PD98059) significantly reduced DEA-NO and 8-Br-cGMP promoter activation and mRNA expression of MMP-13 in transfected BAEC. Moreover, a complex between PKG1-␣ and the G-protein Raf-1, an upstream activator of the extracellular signal-regulated kinase signaling pathway, was detected in cells overexpressing PKG1-␣ or treated either with DEA-NO or 8-Br-cGMP. Thus, we propose that the NO-cGMP-PKG pathway enhances MMP-13 expression by the activation of ERK 1,2. This effect of NO may be the result of pathophysiological importance in the context of inflammation or atherogenesis.

Cbfa-1 mediates nitric oxide regulation of MMP-13 in osteoblasts

“…Involvement of the NO-cGMP in the activation of Cbfa-1-mediated gene expression has been reported in the case of genistein (Pan et al, 2005). The results shown here, as well as in endothelial cells (Zaragoza et al, 2002a;Zaragoza et al, 2002b), identify cGMP as a mediator of NO-induced expression of MMP-13. In this setting, phosphorylation of Cbfa-1 by PKG might have an important role.…”

“…However, the kinase(s) responsible for NO-mediated Cbfa-1 activation have not been identified yet. In endothelial cells, MMP-13 expression is regulated by NO through the cGMP-PKG and ERK pathways (Zaragoza et al, 2002b). Taking into account the data shown by others Pan et al, 2005), and the fact that overexpression of PKG stimulates both the activity of the MMP-13 promoter and MMP-13 expression, we infer that, in osteoblasts, PKG is also able to phosphorylate Cbfa-1.…”

“…3B). In addition, overexpression of the PKG1-␣ dominant-positive isoform (Zaragoza et al, 2002b) also stimulated MMP-13 promoter activity and MMP-13 expression (Fig. 3C).…”

“…Plasmid pMMP-13 OSE-2 is identical to pMMP-13 WT except for a mutation at the OSE-2 responsive element (Pendas et al, 1997). Dominantpositive PKG, containing the catalytic subunit of cGMP-dependent protein kinase, was expressed in MC3T3-E1 cells as described (Zaragoza et al, 2002b). Recombinant Cbfa-1 was cloned by inserting Cbfa-1 cDNA into the SphI and SalI restriction sites of the pQ30 vector (Qiagen).…”

A Neuronal Isoform of Protein Kinase G Couples Mitogen-Activated Protein Kinase Nuclear Import to Axotomy-Induced Long-Term Hyperexcitability inAplysiaSensory Neurons

“…The activation of both apPKG and ap-MAPK by injury was intriguing, because it suggested a possible link between these two kinases (Zaragoza et al, 2002). Indeed, when we added active apPKG to a neuronal lysate, it not only activated ap-MAPK (Fig.…”